TransGenex Nanobiotech (TGN) Inc proposes to further engineer and investigate the potential of a novel fiber inspired smart scaffold to expand cancer stem cells (CSCs), the FiSSCSC platform, for commercialization. CSCs comprising about <1% of cells in tumors escape conventional cancer therapies due to their drug resistance capability and cause cancer relapse and hence expanding CSCs remain an urgent unmet need. TGN is commercializing 3D FiSSTM platform that promotes aggregation of tumor cells resulting in tumoroids. TGN/USF team recently discovered that the multicell (co-cultures of tumor cells with fibroblast and endothelial cells) tumoroids on FiSS significantly expand CSCs, which led to the hypothesis that increasing the robustness of tumoroids by incorporating immature myeloid cells and further engineering of FiSS to maximize mechanical, chemical and biological cues will significantly expand %CSCs in tumoroids. To test this, Aim#1 proposes to expand and characterize MCF7-CSCs and test the potential of immature myeloid cells and other mechanical, chemical and biological cues for CSC expansion. Aim#2 will focus on CSC isolation from in vivo tumors, scale-up, and storage. This highly innovative FiSSCSC platform is expected not only to produce 10 million CSCs/batch, but also increase their long-term storage and stable CSC phenotype for cancer research.